MXPA02002265A

MXPA02002265A - Enantiomerically enriched compounds having photocleavable bond(s) and methods related thereto.

Info

Publication number: MXPA02002265A
Application number: MXPA02002265A
Authority: MX
Inventors: M Skead Benjamin
Original assignee: Qiagen Genomics Inc
Priority date: 1999-08-31
Filing date: 2000-08-31
Publication date: 2003-04-10
Also published as: WO2001016090A1; CN1371356A; KR20020037042A; JP2003508378A; CA2384191A1; AU7349000A; BR0013024A; EP1208079A1

Abstract

The present invention provides a compound having the formula (I) or (II): wherein R1 is selected from halogen and organic moieties; R2 and R3 are independently selected from hydrogen and organic moieties having a mass greater than 15 Daltons where R2 and R3 can together form a carbonyl group or may be joined together within a cyclic structure; Z is an (nplus;1) valent atom excluding carbon where n is an integer greater than 0; R4 is independently selected from hydrogen, halogen, and organic moieties having a mass greater than 15 Daltons, with the proviso that at least one R4 (namely R4a) is an organic moiety having a mass greater than 100 Daltons; R5 at each occurrence is independently either halogen or an organic moiety having a mass of less than 500 Daltons; m is selected from 0, 1, 2, 3 and 4; wherein if R2 = R3 = H, then R1 is not CO2(H r CH3) when Z(R4)n is either of NH(CO) CH(iBu) NH(CO) (CH2Ph or O t Bu), and R4 is not CH2CO2t Bu when Z is OH; and wherein if compounds of both formulae (I) and (II) are present in an admixture, the molar formula (I):formula (II) ratio in the admixture is other than 50: 50. The compounds are useful as tags, including tags detectable by mass spectrometry.

Description

ENATIOMERICALLY ENRICHED COMPOUNDS THAT HAVE LINK (S) FOTODESDODOBLABLE (S) AND METHODS RELATED TO THEM The present invention relates generally to tr 'enantiomerically enriched compounds, specifically enatiomerically enriched compounds having one or more unfolding bonds, as well as methods relating thereto, including synthetic methods.

BACKGROUND OF THE INVENTION Although the labeling of fluorescence and radiolabelling of DNA is widely used for detection purposes in molecular biology and genetics, the number of labels is too small for **? the high-level multiplexing and is not suitable for the rapid acquisition of data, on a large scale. A new class of labels, comprised of small molecule, double-mass spectrometry labels (CMSTs), has been developed and used in applications of gene expression measurement and single nucleotide polymorphism (SNP) genotyping (see, for example, one aspect of PCT International Publication Nos. WO 99/05319, WO 97/27331, WO 97/27327 and WO 97/27325. The system is based on the covalent attachment of CMSTs to the oligonucleotides with a linker, such as linker photodedoblable.Each label has a different mass that is specific to a designated oligonucleotide sequence.The identification of the alleles or Present expressed sequences are determined, for example, by photolytic cleavage of the oligonucleotides and simultaneous detection with a standard single quadruple mass spectrometer using chemical ionization of atmospheric pressure (APCl). The present invention provides advantageous labels, including labels detectable by mass spectrometry, and methods of their use, as more fully described herein.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides enantiomerically enriched compounds having photo-double bonds, and methods for preparing molecules labeled with enantiomerically enriched photodegradable labels. The present invention provides advantages including the ability to obtain a unique distinct product in those cases where there is a need to isolate the members of a plurality of structurally similar tagged molecules, as well as additional related advantages as described more fully. complete in the present. In one aspect, the present invention provides a compound having the formula (I) b (II): (I) (p) wherein R1 is selected from halogen and organic portions; R2 and R3 are independently selected from hydrogen and organic portions having a mass greater than 15 Daltons, wherein R2 and R3 together can form a carbonyl group or can be joined together within a cyclic structure: Z is a valent atom (n + 1 ) that excludes carbon where n is an integer greater than 0; R4 is independently selected from organic portions, hydrogen and halogen having a mass greater than 15 Daltones, with provision that at least one of R4 (mainly R4a) is an organic portion having a mass greater than 100 Daltones; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; where if R2 = R3 = H, then R1 is not CO2 (H or CH3) where Z (R4) n is either -NH (CO) -CH (Bu) -NH (CO) - (CH2Ph or Of-Bu), and R1 is not CH2CO2i-Bu when Z is OH; and wherein if the compounds of both formulas (I) and (II) are present in a mixture; the molar ratio of the formula (I): formula (II) in the mixture is different from 50:50. < In various embodiments of the invention, Z is nitrogen; R4a is detectable by mass spectrometry; R2 and R3 are each hydrogen; R1 comprises synthetic or natural biological material (eg, nucleic acid, protein or saccharide); and / or R4a has a mass less than 10,000 Daltons and a molecular formula of C1.500N0-100O0-100S0 -? oPo-? oHaFßld where the sum of a, ß and d is sufficient to satisfy the otherwise unmeasured valencies of the atoms of C, N, O, P and S. In still another modality, R4a has the formula T2- (J-T3-) P-; wherein T2 is an organic portion formed of carbon and one or more of hydrogen, fluoride, iodine, oxygen, nitrogen, sulfur and phosphorus; which has a mass of 15 to 500 Daltones. T3 is an organic portion formed of carbon and one or more of hydrogen, fluoride, iodine, oxygen, nitrogen, sulfur and phosphorus, having a mass of 50 to 1000 Daltons; J is a direct bond or functional group selected from amide, ester, amine, sulfur, ether, thioester, disulfide, thioether, urea, thiourea, carbamate, thiocarbamate, Schiff's base, reduced Schiff base, mine, oxime, hydrazone, phosphate, phosphonate, phosphoramide, phosphonamide, sulfonate, sulfonamide or carbon-carbon bond; p is an integer that varies from 1 to 50, and when n is greater than 1, each T3 and J are independently selected. In another embodiment, R4a has a formula comprising: wherein G is (CH2)? - ß wherein a hydrogen in one and only one of the CH2 groups of each G is replaced with - (CH2) w- Amida-T4; T2 and T4 are organic portions of the formula C? -2d 0-9? 0-9So -3Po-3HaFpld where the sum of a, ß and d is sufficient to satisfy the otherwise unfilled valences of the C atoms, N, O, S and P. OO li 11 -NC- - CN- ¿10 ¿10 Amide is * "; R10 is hydrogen or C1.10 alkyl, is an integer ranging from 0 to 4, and n is a integer ranging from 1 to 50 so that when n is greater than 1, G, c, Amide, R1 and T4 are independently selected The invention further provides a composition comprising compounds of the formulas (I) and / or ( II) wherein the molar ratio of the formula (I): formula (II) in the compositions are within the range of 95: 5 to 100 or within the range of 5:95 to 0: 100. The invention also provides a process * to provide an enantiomerically enriched compound which comprises contacting a compound of the formula: wherein Z is selected from oxygen, nitrogen and sulfur; wherein R6 is hydrogen when Z is oxygen or sulfur, and when Z is nitrogen then R6 is selected from hydrogen and C? -C22 hydrocarbon and two R6 groups are attached to Z; R5 in each occurrence is independently either halogen or an organic portion that has a mass less than 500 Daltones, m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion, and the bond represented by both a striped and solid line represents either a double or single bond; with an agent selected from (a) an enzyme; (b) an enzyme and an adjuvant of the formula H2N-C (= O) -CHR8-NH-R9, wherein R8 is an organic portion and R9 is an amino protecting group; (c) a chiral acid; (d) a chiral amine; (e) hydrogen and a chiral hydrogenation catalyst; and (f) a mechanical glass separation means; to provide an enantiomerically enriched compound of the formula: In a modality of the process identified above, the compound to be activated in the process has the formula: wherein R in each occurrence is independently either halogen or an organic portion having a mass of less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; and the compound is contacted with an enzyme to provide an enantiomerically enriched compound of the formula: In another embodiment of the process identified above, the compound to be activated in the process has the formula: wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion, and the compound is contacted with an enzyme and an adjuvant of the formula H2N-C (= O) -CHR8-NH-R9, wherein R8 is an organic portion and R9 is an amino protecting group; to provide an enantiomerically enriched compound of the formula: In another modality of the process identified above, the compound to be triggered in the process has the formula: wherein R8 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; and the compound is contacted with a chiral acid, to provide an enantiomerically enriched salt of the formula: Chiral In another modality of the process identified above, the compound to be activated in the process has the formula: wherein R in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltons; m is selected from 0, 1, 2, 3 and 4; and the compound is contacted with a chiral amine, to provide an enantiomerically enriched salt of the formula: Amine In another embodiment of the process identified above, the compound to be activated in the process has the formula: wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; and the compound is contacted with hydrogen in the presence of a chiral hydrogenation catalyst, to provide an enantiomerically enriched compound of the formula: In another aspect, the invention provides compounds of the formula (V) (VI) wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; Z is SH; or NR8 wherein R8 is either hydrogen or an amine protecting group; and wherein if the compounds of both formula (V) and (VI) are present in a mixture, the molar ratio of formula (V): formula (VI) in the mixture is different to 50:50. In another aspect, the invention provides compounds of the formula (VII) (vpi) wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R11 is a functional moiety that includes a phosphoramidite or portion of H-phosphonate; R12 is an organic portion that has a mass of 15-10,000 Daltones; and where if the compounds of both formula (VII) and (VIII) are present in a mixture, the molar ratio of the formula (Vll): formula (HIV) in the mixture is different from 50:50. In one embodiment, in the compounds described above, the phosphoramidite portion of R1 has the formula -OP (OR13) (N (R14) 2) wherein each of R13 and R14 is independently selected from an alkyl group or an alkyl group substituted having one or more substituents selected from halogen and cyano; and two R14 groups can be joined together to form a heterocycloalkyl group with the nitrogen of the phosphoramidite. In another embodiment, the H-phosphonate portion of R11 comprises the formula -O-P (= O) (H) (O- + HN (R15) 3) and R15 is independently an alkyl group These and other aspects and embodiments of the present invention will be apparent from the reference to the following detailed description. For this purpose, several references are set forth herein that describe in more detail certain procedures, compounds and / or compositions, and are incorporated herein by reference in their entirety. DETAILED DESCRIPTION OF THE INVENTION The present invention provides labels suitable for coupling to nucleic acids or other molecules of interest. When coupled to DNA, labels provide a means to achieve a high-throughput genotyping system. Preferably, the linker between the label and the molecule of interest should be completely split under conditions that do not cause fragmentation of the label. In addition, the label preferably produces a maximum value by injected oligonucleotide, and gives optimal signal in terms of ion current. Also, the tag is preferably stable to PCR, HPLC, and other manipulations used in analytical formats.

The reagents to introduce labels to a molecule of interest must proceed reproducibly and in good production. In a preferred approach to meet these objectives, a modular structure was developed, which allows the CMST components to be perfected independently. The components of a CMST preferably include a photolabile linker, a mass spectrometry sensitivity enhancer (MSSE), and a variable mass unit (VMU), all connected together through a scaffold. The present invention provides tagged molecules of interest, precursors for tagged molecules of interest, and methods for generating the tagged molecules and precursors thereto, wherein the tag is enantiomerically enriched. By using enantiomerically enriched precursors, purification of the reaction product of the precursor and the molecule of interest, in order to provide purified labeled molecules of interest, is facilitated. In this manner, the present invention provides a compound having the formula (I) or (II): a) wherein R1 is selected from halogen and organic portions; R2 and R3 are independently selected from hydrogen and organic portions having a mass greater than 15 Daltons, wherein R2 and R3 together may form a carbonyl group or may be linked together within a cyclic structure: Z is a valent atom (n + 1 ) that excludes carbon where n is an integer greater than 0; R4 is independently selected from organic portions, hydrogen and halogen having a mass greater than 15 Daltons, with the proviso that at least one of R4 is an organic portion having a mass greater than 1 00 Daltones; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4. The compound of the invention is enantiomerically enriched, so that if the compounds of both formulas (I) and (II) are present in a mixture; the molar ratio of the formula (I): formula (II) in the mixture is different from 50:50. The compound of the invention includes a 2-nitrophenyl group, in the 1-position of the phenyl ring, a substituted methyl group, wherein the carbon atom of the methyl group will be referred to herein as either C1 or the benzyl carbon atom . In addition to the 2-nitrophenyl group, the C1 atom is directly attached to a hydrogen atom (not shown), a carbon atom (referred to herein as C2) and a carbonless atom. By virtue of the 2-nitrophenyl group, the compounds of the invention are photo-bent. That is, the compounds of the invention will respond to contact with certain electromagnetic radiation by undergoing a cleavage reaction whereby the substituted Z atom is separated from C1 as shown in the following structures, where "^ y ^ f indicates a bond which is photolytically unstable.

Because C1 is directly attached to only one atom that is not a hydrogen or carbon atom, the compounds of the invention undergo a selective photo-unfolding reaction. That is, the C1-Z bond can be selectively split by appropriate photolytic conditions, leaving the other C1 bonds long, if not completely intact. This selective unfolding can occur as long as Z is not a carbon atom. In preferred embodiments, Z is selected from oxygen, nitrogen and sulfur. Each one of oxygen, nitrogen and sulfur is a preferred atom for the Z position. The photolytic conditions that allow the unfolding selective of the bond C1-Z sdn greatly unaffected by another substitution in the 2-nitrophenyl ring. Accordingly, any one or more of the hydrogen atoms of the phenyl ring can be replaced with an equal number of groups Rd. Accordingly, the compounds of the invention can have a number Cm ") of groups R5, wherein m is a selected integer of 0, 1, 2, 3 and 4. The desired photo-duplexing chemistry is greatly independent of the nature of the group Rd, so that R5 can be an inorganic group, eg, halogen, or an organic group Typically, an organic R5 group is not too long, and according to the above has less than about 500 Daltons.The preferred R5 groups are selected from C? -C22 hydrocarbon groups In the preferred compounds of the invention , m is equal to zero, so that the 2-nitrophenyl ring has four hydrogen substituents.After experiencing a photo-doubling reaction, the resulting Z-containing residue that is derived from a computer this of the invention will be referred to herein as a label. The label is, or includes, a detectable portion, wherein this term refers to a portion that can detect, and preferably be characterized, by an analytical method. Representative analytical methods include potentiometric or spectro-metric methods. Representative spectrometric methods include mass spectrometry, infrared spectrometry, ultraviolet spectrometry and fluorescent spectrometry. A representative pontenciometric method is potentiostatic amperometry. In order to provide an effective label, at least one group R4 is an organic portion having a mass greater than 100 Daltones. This group R4 will be referred to herein as the first group R4 (or R4a). The fact that labels should have masses greater than 100 Daltones is desirable for a number of reasons. When the labels have masses less than 100 Daltones, it can be difficult to identify the labels. For example, when labels are identified and characterized by mass spectrometry, labels of molecular weight less than 100 Daltons can be difficult to distinguish from background noise inherent in the operation of a mass spectrometer. In addition, the present invention provides compositions containing a number of non-identical compounds of the invention, wherein the non-identical compounds contain non-identical labels that can be distinguished from each other by an analytical method. In order to distinguish a number of non-identical labels, the labels must be sufficiently different from each other that the analytical method can detect the difference. In order to obtain an adequate number of labels that are distinct by, for example, potentiometry, the labels must contain a number of atoms that can be joined together in non-identical ways that are analytically distinct. When labels have a mass of less than 1 00 Daltones, there is an unduly limiting number of analytically distinct labels. In accordance with the above, to allow sufficient variability in structure, and to allow easy and efficient detection, Labels must have masses in excess of 100 Daltones. Provided the first group R4 has a mass in excess of 100 Daltons, the identity of the other group R4 in a compound of the invention can be selected to achieve one or more goals. For example, a second or more R4 groups (e.g., R4b, R4c, etc.) can impart information that complements the information derived from the first R4 group. In this case, a second or more groups R4 also preferably has a mass of at least 100 Daltones. However, in other cases, it may be desirable that the second or more R4 groups no longer satisfy the otherwise dissatisfied valence state of C1, do not interfere with the photo-double response reaction or detection of the label, and may be synthetically easy to to form. In the latter case, the second or more R4 groups may be hydrogen or simple hydrocarbon groups. In a preferred embodiment, the second or more groups R4, if present, is / are hydrogen. In the compounds of the invention, R1 is selected from halogen and organic portions, while R2 and R3 are independently selected from hydrogen and organic portions having a mass greater than 15 Daltones. As stated above, the compounds of the invention include a detectable moiety wherein detection, and optionally characterization of the detectable moiety provides information about the compound of the invention. In one embodiment, the group R1 contains the characteristic of the compound about which the information is desired. For example, R1 may include biological material from an individual, and the label is characteristic of that individual and / or the material specific biological As another example, the R1 group may contain a particular synthetic nucleic acid, peptide or saccharide sequence, which is uniquely identified by the label. In another embodiment, R1 is a reactive chemical group that can undergo a chemical reaction whereby the biologically derived material can be photo-degradablely linked to a label. For example, R1 can be a halogen atom that can be displaced in a chemical reaction by biological material or derived from biological material. The groups R2 and R3 are independently selected from hydrogen and organic portions having a mass greater than 15 Daltones. Typically, R2 and R3 are selected with a view to their impact on the chemistry selected to bind R1 to C1, and / or for their impact on the photo-double reaction whereby the label is separated from C1. When each of R2 and R3 is hydrogen, then neither R2 * nor R3 adversely impacts the photo-double reaction, and are typically compatible with any chemistry used to couple R1 to C1. According to the foregoing, R2 and R3 are preferably hydrogen. However, R2 and R3 could be groups other than hydrogen. For example, R2 and R3 together can form a carbonyl group with C1. Alternatively, R1 and R2 can, together with C1, form a cyclic structure. As another alternative, either R2 or R3 may be an organic portion of mass greater than 15 Daltones, for example, a C? -C22 hydrocarbon group. Typically, the hydrocarbon groups do not interfere with the desired photo-unfolding reaction, and do not interfere with the coupling chemistry R1 / C1.

Another characteristic of a compound of the invention is that the C1 atom is chiral. As seen from the formulas (I) and (II), the nature of the substituents in C1 necessarily requires C1 to be qu? Ral. Furthermore, if the compounds of the formulas (I) and (II) are in mixture, the molar ratio of the compounds of the formula (I) to the compounds of the formula (II) is different to 50:50. In a preferred embodiment of the invention, the compounds of the formula (I) are not in admixture with the compounds of the formula (II), and vice versa. Furthermore, in the case where the compounds of the formula (I) are in mixture with the compounds of the formula (II), the molar ratio of the compounds of the formula (I) to the compounds of the formula (II) it is preferably in excess of 95: 5, or 96: 4, or 97: 3, or 98: 2 or 99: 1, or in excess of 99.5: 0.5, or vice versa (i.e., the molar ratio of the compounds of the formula (II) to the compounds of the formula (I) is preferably in excess of 95: 5, or 96: 4, or 97: 3, or 98: 2 or 99: 1, or in excess of 99.5: 0.5) . Because the compounds of the formulas (l) or (II) are either found or not, or only minimally are found, in admixture with the compounds of the formulas (II) or (I), respectively, the chromatographic separations which include the compounds of the formula (!) or (II) are much easier and effective than if significant amounts of the compounds of the formulas (I) and (II) were in mixture with each other. The present invention provides compositions containing a plurality of compounds of the formulas (I) or (li) which, due to their isomeric constitution, can be more easily separated from each other that would otherwise be the case. This increased ability to distinguish labeled compounds in a different manner allows the formation of compositions that could otherwise not be characterized. For example, although two compounds of the formula (I) may be different by virtue of slightly different R 1 groups, these two compounds (compounds IA and IB) may not be effectively separable if they are mixed with compounds of the formula (II) having the identical substitution around C1 (compounds HA and HB). This is because the chromatography elution time of compound IA can overlap that of compound IIB and / or the elution time of compound IB can overlap that of compound HA. According to the above, the compounds IA and IIB may not be easily resolved with each other, due to the interference of the compounds HA and HB. The present invention addresses this problem by providing compounds of the formula (I) free of, or effectively free of, compounds of the formula (II). . In another aspect, the present invention is directed to a process for providing an enantiomerically enriched compound using a specific agent. More specifically, the process acts on a compound of the formula: wherein Z is selected from oxygen, nitrogen and sulfur; where R6 is hydrogen when Z is oxygen or sulfur, and when Z is nitrogen then Rβ is selected from hydrogen and CÍ-022 hydrocarbon and two Rβ groups are attached to Z; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltons, m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic moiety, and the bond represented by both a striped and solid line represents either a double or single bond. The inventive process contacts a compound as identified above with an agent. The agent is selected from (a) an enzyme; (b) an enzyme and an adjuvant of the formula H2N-C (= O) -CHR8-NH-R9, wherein R8 is an organic portion and R9 is an amino protecting group; (c) a chiral acid; (d) a chiral amine; (e) hydrogen and - a chiral hydrogenation catalyst; and (f) a mechanical glass separation means. The compound is contacted with the agent to provide an enantiomerically enriched compound of the formula In addition to the specific techniques described herein for preparing compounds that have photolabile links to enantiomerically enriched labels, other suitable techniques can be found in the following references: Asymmetric Catalysis in Organic Synthesis, R. Noyori, John Wiley & Sons, New York, NY, 1994; Asymmetric Synthetic Methodology, D, J. Agér and M. B. East, CRC Press, Boca Raton, FL, 1995; Asymmetric Synthesis, R.A. Aitken and S.N. Kilenyi, Eds. Blackie Academic & Professional, Glasglow, R.U., 1992; Asymmetric Synthesis, J. Morrison,. Ed., Academic Press, Orlando, FL. (series, including volumes issued in 1984 and 1985); Asymmetric Synthesis, R.G. Proctor, Oxford University Press, New York, NH, 1996; Asymmetric Synthesis: Construction of Chiral Molecules using Amino Acids, G.M. Coppola and H.F. Schuster, John Wiley & Sons, New York, NY, 1987; Catalytic Asymmetric Synthesis, I. Ojima, Ed., VHC Publishers, New York, NY, 1993; Chiral Auxiliaries and Ligands in Asymmetric Synthesis, J. Seyden-Penne, John Wiley & Sons, New York, NY, 1995; Chiral Separations; Applications and Technology, S., Ahuja, Ed., American Chemical Society, Washington, D.C. , nineteen ninety six; Chirality in Industry l & II, A.N. Collins, G.N. Sheldrake and J. Crosby, Eds. , John Wiley & Sons, New York, NY, 1995 and 1997; Chirotechnologu: Industrial Synthesis of Optically Active Compunds, R.A. Sheldon, Marcel Dekkar, New York NY 1993; and Enantiomers, Racemates and Resolutions, J. Jacques, A. Collet, and S.H. Wilen, John Wiley & Sons, New York, NY, 1981. The present compounds and compositions, and methods for their preparation, can be used to provide target molecules wherein the label is both optimally enriched and photodedobly linked to the molecule. Molecules with photo-detachably linked labels, which may be provided in optically enriched form according to the present invention, as well as methods that may be practiced with compounds and compositions of the present invention, are set forth in, for example, International PCT Publications Nos. WO 99/05319; WO 97/27331; WO 97/27327; WO 97/27325; and WO 95/04160. In this manner, the compounds of the present invention can be substituted for the labeled compounds described in these four publications, as well as other methods and analyzes wherein molecules having photobledably linked labels are used. The following examples are established as a means to illustrate the present invention and are not constructed as a limitation thereto. EXAMPLES In the following examples, and unless otherwise noted, the chemical reagents and reagents were of the standard commercial grade, obtained from commercial supply houses such as Aldrich (Milwaukee, Wl; www.sigma-aldrich.com), Fluka (a division of Aldrich) and Lancaster Synthesis, Inc. (Windham, NH; http://www.lancaster.co.uk). EXAMPLE 1 RESOLUTION WITH QUITAL ACID DTTA The methyl ester of (+) - 3-amino-3- (2-nitrophenol) propionic acid (24.6 g, 10 mmol) and the di-p-toluoyl-D-tartaric acid of Chiral acid (DTTA, 42.4 g, 10 mmol) was taken in 500 mL of methanol on heating at 67 ° C. The resulting solution was cooled to 56 ° C with the methyl ester salt of 3-amino-3- (2-nitrophenyl) propionic acid and DTTA. The mixture was allowed to cool to room temperature (21 ° C) and stirred for 16 hours. The crystals that formed were collected by filtration giving a white solid that is enriched in an enantiomer (46% enantiomeric excess, e.e.). A crystallization in methanol resulted in 12 g of material with a diastereomeric purity of 88%. EXAMPLE 2 RESOLUTION WITH CANHANIC ACID (-) OF CHAIRAL ACID The (+) - 3-amino-3- (2-nitrophenyl) propionic acid (172 Ng, 0.82 mmol) and the (-) - canfánico of chiral acid (165 mg) 0.83 mmol, Fluka) were taken up in 5 mL of methanol on heating to reflux. The resulting solution was cooled to room temperature (21 ° C). The crystals that formed were collected by filtration to give a white solid (105 mg, 31% of production). In analysis by chiral high pressure liquid chromatography (cHPLC, Chirobiotic T column at 5 ° C, 75% regulator of 20 mM ammonium acetate, pH 4.5 and 25% EtOH, detector at 215 nm) indicated that the product it was enantiomerically pure. EXAMPLE 3 SELECTIVE HYDROLYSIS WITH ENZYME The methyl ester of (+) - 3-amino-3- (2-nitrophenyl) propionic acid (60 g, 270 mmol) was dissolved in phosphate buffer (50 mM, 500 mL) and adjusted to pH 7. After the Enzyme Amano PS (6 g, 10% by weight Enzyme Amano (Milton Keynes, UK; Lombard IL, USA; www.amano-enzyme.co.hp)) as a mixture in phosphate buffer (100 mL) was added to the solution, and the reaction mixture was stirred at room temperature (21 ° C) for 24 hours. At the termination, the amino acid precipitate was filtered and the aqueous filtrate was removed before the solid was rinsed with dichloromethane (DCM, 2 x 10 mL). The solid is dissolved in aqueous HCl (6 M, 150 mL) but a fine unknown precipitate, which was filtered, still remains. The resulting aqueous mixture is extracted with DCM before the solid §e will precipitate by adjusting the pH to 7. This gives the desired amino acid (23 g, 90% production). The initial filtrate was basified to pH 8 and extracted first with the DCM washes collected above; and then with fresh DCM. The organic layers were combined, dried and evaporated to give the ethyl ester amino (23 g, 80% yield). Optical rotation of the ethyl ester HCl salt in methanol gave an aD of 127.55 ° (c = 1.20 ° C, 589 nm). The enantiomeric excess (e.e.) of both the amino acid and the ethyl ester amino was determined by cHPLC and both were in excess of 98% enatiomerically pure. * EXAMPLE 4 SYNTHESIS OF REACTIVE MOLECULE TO SUPPLY LABEL OPTICALLY ENRICHED TO THE MOLECULE OF INTEREST * The labels described herein are preferably conjugated with a molecule of interest through a photolabile bond. A photolabile linkage occurs within an o-nitrobenzyl group (see, for example, (a) Greenberg, M.M .; Gilmore, J.L. J. Org.

Chem. 1994, 59, 746-753; (b) Yoo, D.J; Greenberg, M.M. J. Org. Chem. 1995, 60, 3358-3364; and (c) Venkatesan, H .; Greenberg, M.M. J. Org. Chem 1996, 61, 525-529). The reaction of intramolecular photoredox in an o-nitrobenzyl group (see, for example, Pillai, V.N.R. Synthesis 1980, 1-26) allows the label to be rapidly split from the oligonucleotide under neutral conditions (six seconds of exposure with one lamp). 254 nm Hg) and without fragmentation. In accordance with the above, the synthetic route begins (Scheme 1) with esterification of the photosensitive eniazador of 3-amino-3- (2-nitrophenyl) propionic acid, (ANP, i, see, for example, Brown, BB: Wagner, DS, Geysen, HM, Molecular Diversity 1995, 1, 4-12, and (b) Rodebaugh, R .; Fraser-Reid; B .; Geysen, HM Tetrahedron Letters 1997, 38, 7653-7656) to give ethyl ester hydrochloride (2) in 84% of production. This is followed by the enzymatic transformation of 2 to provide the ethyl ester as a single isomer, which facilitates the purification of HPLC in the oligonucleotide conjugation step. The ethyl ester hydrochloride is dissolved in water and pH neutral is adjusted to pH with 2N HCl. The PS Amano enzyme is added to a phosphate buffer mixture. After completion of the reaction, a basic lift removed the hydrolysed ANP byproduct (4), and the single isomer ethyl ester (3,> 99% e.e.) was recovered (92% of available material). Scheme 1 Lysine provided scaffolding that was very suitable for the modular approach to the synthesis of labels. The two carboxylic acid and amine groups present in the lysine provided for robust peptide chemistries with resultant amide bonds between the tag components. Peptides have a predictable behavior in mass spectrometry and are also relatively stable at 254 nm of light. As seen in Scheme 2, the coupling of 3 with a-BOC-e-Alloc-lysine (5), using EDAC1 (1 - [3- (Dimethylamino) propyl] -3-ethyl-carbodiimide hydrochloride, obtained from Aldrich Chemical Co., Milwaukee, WI) and 1-hydroxybenzotriazole (HOBT), gave the protected ANP lysine (6). Removal of BOC with HCl provided the e-Alloc-lysine ester (7) as a white solid in 81% yield of 3. Perfluorinated aromatics can be used as an electrophoretic label for analytical purposes in mass spectrometry in a negative ion mode ( see, for example, (a) Abdel-Baky, S .; Klempier, N .; Giese, RW Tetrahedron 1990, 46, 5859; (b) Abdel-Baky, S .; Allam, K .; Giese, RW Anal. C jem.1993, 65, 498-499; (c) Trainor, T; Giese, RW, Vouros, P. Journal of Chromatography 1988, 452, 369-376; and (d) Saha, M.; Saha, J; Giese, RW Journal of Chromatography 1993, 641, 400-404). Our initial strategy for an MSSE candidate, the ionization component of the label, consists of linking several highly fluorinated carboxylic acids, deficient in electron to a representative scaffold and measure the relative ionic current with APCl in a negative ion mode. Fragmentation occurred, giving multiple maximum values together with a low signal. The boosters for the opposite ionization regime were then explored. Labels were made incorporating pyridyl, prolinyl and piperidinyl structures as MSSE and tested in APCl in a positive way. By all criteria, a label that uses N-methyl isonipecotic acid (INA) gives excellent results. The INA hydrochloride was coupled to 7, using EDAC and triethylamine, to give crude Alloc-protected structure 8, which was deprotected with diethylamine, triphenylphosphine and palladium acetate. The resulting core structure (9) was crystallized from the reaction mixture and recovered by filtration in 95% yield as a yellow solid. Scheme 2 To provide a long set of detectable labels with different molecular weights, the core structure 9 was derived with a set of carboxylic acids referred to as variable mass units (VMUs). The masses of the VMUs were spaced at a minimum of 4 a.m.u. apart, to minimize the isotope overlap between the labels. The molecular weights of the VMUs varied from 90-298 a.m.u. Preferred VMUs do not have the following properties or characteristics: (1) functionalities incompatible with the synthetic sequence, such as esters; (2) elements with multiple isotopes (Cl, Br, S); (3) functionalities that can lead to competing photoprocesses (iodides, acyl and aryl-phenones); (4) racemic acids; and (5) lack of commercial availability VMUs were coupled to 9, using HATU and N-methyl morpholine. After purification by column chromatography, the ethyl ester CMST (10) was recovered in variable yields. For the final step, formation of the active ester was achieved using a catalysed base transesterification of tetrafluorophenyl trifluoroacetate (TFP TFA, see, for example, Green, M .; Berman, J. Tetrahedron Letters 1990, 31, 5851-5852; TFP TFA was prepared in a manner similar to pentafluorophenyl trifluoroacetate, and was used because the proton in the tetrafluorophenyl ring acts as a diagnostic NMR check) and 11, which results in the TFP ester CMST (12) in variable yields. The active tetraflurophenyl ester was chosen because of the easily removed side products and the compatibility and relative stability towards the oligonucleotide conjugation conditions. The TFP CMST esters were conjugated with 5'-aminohexyl back oligonucleotides (obtained from TriLink Biotechnology, San Diego, CA) according to the method of Lukhtanov et al., (See, for example, Lukhtanov, EA; Kutyavin, IV, Gamper, HB; Meyer, RB Jr. Bioconjug, Chem. 1995, 4, 418-426). Scheme 3 The number of labels was also expanded by the incremental use of a tyrosine derivative that functioned as a coarse mass adjuster (GMA, and more specifically N-Boc-O-ethyl-L-tyrosine, which can be commercially ordered from Bachem California ( Torrance, CA) or prepared by alkylation of N-Boc-L-Tyrosine-Ome with iodoethane and cesium carbonate in 86% yield, followed by hydrolysis with NaOH in quantitative) which increases the molecular weight of the label split by 191 amu and allows the set of VMUs to be re-used for labels of higher molecular weight. GMA was coupled to the core structure 9 as shown in Scheme 4. After the basic survey, the GMA structure protected with BOC was isolated by filtration. Deprotection with TFA resulted in the monomer core structure GMA (13, n = 1) in quantitative production. VMUs are coupled in the same manner as before, resulting in additional sets of labels (14). The use of four units of the mass adjuster, a total of almost 200 labels can be synthesized. Scheme 4a a (a) EDAC, HOBT, THF; (b) TFA, DCM; (c) VMU, HATU, NMM; (d) 1 N NaOH, THF; (e) TFP, TFA, DIEA, DMF From the foregoing, it will be appreciated that, although the specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without departing from the spirit and scope of the invention. In accordance with the foregoing, the invention is not limited except as by the appended claims.

Claims

CLAIMS A compound having the formula (I) or (II): wherein R1 is selected from halogen and organic portions; R2 and R3 are independently selected from hydrogen and organic portions having a mass greater than 15 Daltons, wherein R2 and R3 together may form a carbonyl group or may be linked together within a cyclic structure: Z is a valent atom (n + 1 ) that excludes carbon where, n is an integer greater than 0; R4 is independently selected from organic portions, hydrogen and halogen having a mass greater than 15 Daltones, with the proviso that at least one of R4 (mainly R a) is an organic portion having a mass greater than 100 Daltones; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; where if R2 = R3 = H, then R1 is not CO2 (H or CH3) where Z (R4) n is either -NH (CO) -CH (iBu) -NH (CO) - (CH2Ph or Of -Bu), and R1 is not CH2CO2f-Bu when Z is OH; and wherein if the compounds of both formulas (I) and (II) are present in a mixture; the molar ratio of the formula (I): formula (II) in the mixture is different from 50:50. 2. A compound according to claim 1, characterized in that Z is nitrogen. 3. A compound according to claim 1, characterized in that R4a is detectable by mass spectrometry. 4. A compound according to claim 1, characterized in that R4 has a mass of less than 10,000 Daltons and a molecular formula of C? -5ooNo-? OoOo-? OoSo -? OPo-? OHaFßld where the sum of a, ß and d is sufficient to satisfy the otherwise unfilled valences of the C, N, O, P and S atoms. 5. A compound according to claim 1, characterized in that R4a has the formula T - (J-T3-) P-; wherein T2 is an organic portion formed of carbon and one or more of hydrogen, fluoride, iodine, oxygen, nitrogen, sulfur and phosphorus; which has a mass of 15 to 500 Daltones; T3 is an organic portion formed of carbon and one or more of hydrogen, fluoride, iodine, oxygen, nitrogen, sulfur and phosphorus, having a mass of 50 to 1000 Daltons; J is a direct bond or functional group selected from amide, ester, amine, sulfur, ether, thioester, disulfide, thioether, urea, thiourea, carbamate, thiocarbamate, Schiff base, reduced Schiff base, imine, oxime, hydrazone, phosphate , phosphonate, phosphoramide, phosphonamide, sulfonate, sulfonamide or carbon-carbon bond; and p is an integer ranging from 1 to 50, and when n is greater than 1, each T and J are independently selected. A compound according to claim 1, characterized in that R has a formula comprising: wherein G is (CH2)? _ ß wherein a hydrogen in one and only one of the CH2 groups of each G is replaced with - (CH2) W-Amide-T4; T2 and T4 are organic portions of the formula CL5.5N0-9O0.9S0 -3Po-3HaFpld where the sum of a, ß and d is sufficient to satisfy the otherwise unfilled valences of the C, N, O atoms, S and P: O II -N-C- - C-N- 0 ¿10 Amide is R1 1 .10 R is hydrogen or C- .o alkyl, w is an integer ranging from 0 to 4; and n is an integer ranging from 1 to 50 so that when n is greater than 1, G, c, Amide, R1 and T4 are independently selected. 7. A compound according to claim 1, characterized in that R2 and R3 are each hydrogen. 8. A compound according to claim 1, characterized in that R1 comprises natural or synthetic biological material. 9. A compound according to claim 8, characterized because R1 comprises nucleic acid, protein or saccharide. A composition comprising the compounds of claim 1, characterized in that the molar ratio of formula (I): formula (II) is within the range of 95: 5 to 100: 0 or within the range of 5:95. to 0: 100. 11. A process for providing an enantiomerically enriched compound comprising contacting a compound of the formula: wherein Z is selected from oxygen, nitrogen and sulfur; wherein Rβ is hydrogen when Z is oxygen or sulfur, and when Z is nitrogen then Rβ is selected from hydrogen and C?-C22 hydrocarbon and two Rβ groups are attached to Z; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltons, m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion, and the bond represented by both a striped and solid line represents either a double or single bond; with an agent selected from (a) an enzyme; (b) an enzyme and an adjuvant of the formula H2N-C (= O) -CHR8-NH-R9, wherein R8 is an organic portion and R9 is an amino protecting group; (c) a chiral acid; (d) a chiral amine; (e) hydrogen and a chiral hydrogenation catalyst; and (f) a mechanical glass separation means; to provide an enantiomerically enriched compound of the formula: 12. The process according to claim 1, characterized in that a compound of the formula: wherein R in each occurrence is independently either halogen or an organic portion having a mass of less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; an enzyme is contacted to provide an enantiomerically enriched compound of the formula: 13. The process according to claim 1, characterized in that a compound of the formula: wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; it is contacted with an enzyme and an adjuvant of the formula H2N-C (= O) -CHR8-NH-R9, wherein R8 is an organic portion and R9 is an amino protecting group; to provide an enantiomerically enriched compound of the formula: 14. The process according to claim 11, characterized in that a compound of the formula: '- 3J3 - where R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltons; m is selected from 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; it is contacted with a chiral acid, to provide an enantiomerically enriched salt of the formula: The process according to claim 1, characterized in that a compound of the formula: wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; is contacted with a chiral amine, to provide an enantiomerically enriched salt of the formula: Chiral amine 16. The process according to claim 1, characterized because a compound of the formula: wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; hydrogen is contacted in the presence of a chiral hydrogenation catalyst, to provide an enantiomerically enriched compound of the formula: 17. A compound having the formula (V) or (VI): (V) (VI) wherein R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m - 4p -select 0, 1, 2, 3 and 4; R7 is hydrogen or an organic portion; Z is SH; or NR8 wherein R8 is either hydrogen or an amine protecting group; and wherein if the compounds of both formula (V) and (VI) are present in a mixture, the molar ratio of formula (V): formula (VI) in the mixture is different to 50:50. 18. A compound that has the formula (VII) or (VIII): (VII) (HIV) wherein R5 in each occurrence is independently either halogen or "an organic portion having a mass less than 500 Daltons; m is selected from 0, 1, 2., 3 and 4; R11 is a functional moiety that includes a phosphoramidite or portion of H-phosphonate; R12 is an organic portion that has a mass of 15-10,000 Daltones; and wherein if the compounds of both formula (VII) and (HIV) are present in a mixture, the molar ratio of formula (Vll): formula (HIV) in the mixture is different to 50:50. 19. The compound according to claim 18, characterized in that the phosphoramidite portion of R11 has the formula -O- P (OR13) (N (R14) 2) wherein each of R13 and R14 is independently selected from an alkyl group or a substituted alkyl group having one or more substituents selected from halogen and cyano; and two R14 groups can be joined together to form a heterocycloalkyl group with the nitrogen of the phosphoramidite. The compound according to claim 18, characterized in that the H-phosphonate portion of R11 comprises the formula -OP (= O) (H) (O- + HN (R15) 3) and R1 d is independently an alkyl C group .-H.H. "42" \ r / ~ i "SUMMARY The present invention provides a compound having the formula (I) or (II), wherein R1 is selected from halogen and organic portions: R2 and R3 are independently selected from hydrogen and organic portions which have a mass greater than 15 Daltones, where R2 and R3 together may form a carbonyl group or may be joined together within a cyclic structure: Z is a valent atom (n + 1) that excludes carbon where n is an integer greater than 0; R4 is independently selected from organic portions, hydrogen and halogen having a mass greater than 15 Daltones, with the proviso that at least one of R4 (mainly R a) is an organic portion having a mass greater than 100 Daltones; R5 in each occurrence is independently either halogen or an organic portion having a mass less than 500 Daltones; m is selected from 0, 1, 2, 3 and 4; where if R2 = R3 = H, then R1 is not CO2 (H or CH3) where Z (R4) n is either -NH (CO) -CH (iBu) -NH (CO) - (CH2Ph or Of -Bu), and R4 is not CH2CO2f-Bu when Z is OH; and wherein if the compounds of both formulas (I) and (II) are present in a mixture; the molar ratio of the formula (I): formula (II) in the mixture is different from 50:50. The compounds are useful as labels, including labels detectable by mass spectrometry.